Abstract
Proteins unique to B16 melanoma cells grown in culture and as solid tumors have been independently described recently. As both proteins have apparent Mr of 65,000-70,000, we have more fully compared their biochemical characteristics. The Mr 65,000 protein, which is isolated from B16 cells in culture, has an aggregate molecular weight of >200,000 under nonreducing conditions. This Mr 65,000 form is shed into the culture medium exclusively and is the predominant molecular species of serum-free culture supernatants. It was found that B700, a major membrane protein of B16 cells in solid tumors, and the Mr 65,000 preparation from cultured cells were electrophoretically identical when compared under four separate sets of conditions. These two proteins also share immunologic determinants. The results suggest the identity of these two proteins isolated by completely different approaches.
Keywords: antigens, tumors, polyacrylamide gel electrophoresis, isoelectric focusing
Full text
PDF



Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bhavanandan V. P., Kemper J. G., Bystryn J. C. Purification and partial characterization of a murine melanoma-associated antigen. J Biol Chem. 1980 Jun 10;255(11):5145–5153. [PubMed] [Google Scholar]
- Black P. H. Shedding from the cell surface of normal and cancer cells. Adv Cancer Res. 1980;32:75–199. doi: 10.1016/s0065-230x(08)60361-9. [DOI] [PubMed] [Google Scholar]
- Bystryn J. C., Schenkein I., Baur S., Uhr J. W. Partial isolation and characterization of antigen(s) associated with murine melanoma. J Natl Cancer Inst. 1974 Apr;52(4):1263–1269. doi: 10.1093/jnci/52.4.1263. [DOI] [PubMed] [Google Scholar]
- Embleton M. J., Price M. R., Baldwin R. W. Demonstration and partial purification of common melanoma-associated antigen(s). Eur J Cancer. 1980 Apr;16(4):575–585. doi: 10.1016/0014-2964(80)90238-8. [DOI] [PubMed] [Google Scholar]
- Fidler I. J. Immune stimulation-inhibition of experimental cancer metastasis. Cancer Res. 1974 Mar;34(3):491–498. [PubMed] [Google Scholar]
- Fidler I. J. Selection of successive tumour lines for metastasis. Nat New Biol. 1973 Apr 4;242(118):148–149. doi: 10.1038/newbio242148a0. [DOI] [PubMed] [Google Scholar]
- GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gersten D. M., Marchalonis J. J. A rapid, novel method for the solid-phase derivatization of IgG antibodies for immune-affinity chromatography. J Immunol Methods. 1978;24(3-4):305–309. doi: 10.1016/0022-1759(78)90133-3. [DOI] [PubMed] [Google Scholar]
- Gersten D. M., Marchalonis J. J. Demonstration and isolation of murine melanoma-associated antigenic surface proteins. Biochem Biophys Res Commun. 1979 Oct 12;90(3):1015–1024. doi: 10.1016/0006-291x(79)91928-4. [DOI] [PubMed] [Google Scholar]
- Gupta R. K., Irie R. F., Chee D. O., Kern D. H., Morton D. L. Demonstration of two distinct antigens in spent tissue culture medium of a human malignant melanoma cell line. J Natl Cancer Inst. 1979 Aug;63(2):347–356. [PubMed] [Google Scholar]
- Hearing V. J., Nicholson J. M. Abnormal protein synthesis in malignant melanoma cells. Cancer Biochem Biophys. 1979;4(2):59–68. [PubMed] [Google Scholar]
- Kerney S. E., Montague P. M., Chretien P. B., Nicholson J. M., Ekel T. M., Hearing V. J. Intracellular localization of tumor-associated antigens in murine and human malignant melanoma. Cancer Res. 1977 May;37(5):1519–1524. [PubMed] [Google Scholar]
- Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
- Khosravi M., Preddie E., Hartmann D., Lewis M. G. Human melanoma tumor-specific antigens (II): Comparison of antigens from tumor cell plasma membranes of five melanoma patients. Cancer Biochem Biophys. 1980;4(3):195–202. [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
- McCabe R. P., Ferrone S., Pellegrino M. A., Kern D. H., Holmes E. C., Reisfeld R. A. Purification and immunologic evaluation of human melnoma-associated antigens. J Natl Cancer Inst. 1978 Apr;60(4):773–777. doi: 10.1093/jnci/60.4.773. [DOI] [PubMed] [Google Scholar]
- Righetti P., Drysdale J. W. Isoelectric focusing in polyacrylamide gels. Biochim Biophys Acta. 1971 Apr 27;236(1):17–28. doi: 10.1016/0005-2795(71)90144-9. [DOI] [PubMed] [Google Scholar]
- Ugel A. R., Chrambach A., Rodbard D. Fractionation and characterization of an oligomeric series of bovine keratohyalin by polyacrylamide gel electrophoresis. Anal Biochem. 1971 Oct;43(2):410–426. doi: 10.1016/0003-2697(71)90271-5. [DOI] [PubMed] [Google Scholar]
- Wilson C. M. Studies and critique of Amido Black 10B, Coomassie Blue R, and Fast Green FCF as stains for proteins after polyacrylamide gel electrophoresis. Anal Biochem. 1979 Jul 15;96(2):263–278. doi: 10.1016/0003-2697(79)90581-5. [DOI] [PubMed] [Google Scholar]


